Multicapillary fraction collection system and method

ABSTRACT

A method of injecting, isolating and separating mixed analytes from one or more samples. The method includes collecting successive fractions from each of a plurality of samples at discrete points in time. Fractions may be analyzed at the time of collecting, or later, using one or more detector systems. In one embodiment, a processor controls the elutions of fractions by modulating the migration field in a separation pathway. The processor also controls distribution of the fraction into a particular collection well of a plurality of collection wells.

RELATED APPLICATIONS AND CLAIM OF PRIORITY

[0001] This application claims priority to U.S. Provisional PatentApplication serial No. 60/264,574, entitled METHOD AND APPARATUS FORTESTING SAMPLES UTILIZING A SAMPLING APPARATUS AND ONE OR MORE SEPARATEDETECTORS, and filed on Jan. 26, 2001, the specification of which ishereby incorporated by reference.

[0002] This application claims priority to U.S. Provisional PatentApplication serial No. 60/340802, entitled FRACTION COLLECTION SYSTEMAND METHOD, and filed on Dec. 12, 2001, the specification of which ishereby incorporated by reference.

[0003] This application claims priority to U.S. patent application Ser.No. ______, entitled THIN FILM ELECTROPHORESIS APPARATUS AND METHOD,filed on Jan. 14, 2002, the specification of which is herebyincorporated by reference.

[0004] The specification of U.S. patent application Ser. No. ______,entitled NANOPOROUS MEMBRANE REACTOR FOR MINIATURIZED REACTIONS ANDENHANCED REACTION KINETICS, and filed on Jan. 14, 2002, is herebyincorporated by reference.

TECHNICAL FIELD

[0005] This document relates generally to analysis of samples, includinglarge scale sampling of biological test samples. More specifically, thepresent invention relates to a method and apparatus for analyzingfractions or analytes from a sample.

BACKGROUND OF THE INVENTION

[0006] Large scale testing and analysis is important to many industries,including biotechnology, medical diagnostics, and pharmaceuticals. Forexample, manufacturers in the biotechnology industries implementautomated laboratory systems, such as high throughput processing, totest and analyze large numbers of samples.

[0007] In some cases however, analytical or preparatory techniques arenot suitable for use with automated processing systems. Consequently,certain procedures are performed separately from the automated systemand involve some amount of human intervention, thus increasing theproduction time and cost.

[0008] Capillary electrophoresis (CE), for example, has been used inboth analytical and preparative applications. Among the advantages of CEis the ability to quickly separate similar compounds on a nanoliterscale. For example, CE can be used with mixtures of proteins,macromolecules, nucleotides, enantiomers, and chiral molecules.Pharmaceutical, agricultural, and chemical industries routinely use CEin analytical applications, as well as in research and development.

[0009] The biotechnology industry, for example, has capitalized on theability of CE to quickly analyze small volumes of material. Capillaryelectrophoresis can be used with nucleic acids, separations andanalysis. There remains, however, a need for a rapid process thatidentifies and isolates large volumes of material, such as is generatedby pharmacogenomics and the human genome project.

[0010] Advances in cloning techniques, for example, have enabled thegenomic sequencing of a organisms. A sample of DNA, or a fragmentthereof, from a particular organism, can be cloned and then analyzedusing CE to determine the DNA sequences. Also, CE may be used to isolatea particular DNA fragment for cloning. For example, CE may isolate apreparative amount of a particular DNA fragment from a mixed DNApopulation. This purified fragment can then be inserted into recombinantDNA plasmid, which then clones the corresponding protein.

[0011] Conventional slab gel electrophoresis (SGE) is unsuitable forhigh volume analysis of DNA sequences. Each sample derived from SGE isphysically cut from the slab and separately analyzed, thus requiringhuman intervention. Consequently, these and other disadvantages renderSGE incompatible with an automated, high throughput system.

[0012] Limitations in the speed, volume and efficiency of CE technologyhave impaired efforts to streamline or automate genomic processes. Thus,there remains a need for faster, higher volume, and more efficientmethods of DNA separation, isolation and cloning. In addition, there isa need for an improved system and method for analyzing biological andchemical samples that yields high resolution and rapid results.

SUMMARY

[0013] The present subject matter is directed to apparatuses, systemsand methods for performing high throughput collection of fractions oranalytes. In one embodiment, analysis, or detecting, is integrated intothe present system. In one embodiment, detection is performed as asubsequent process after having collected the various fractions.

[0014] In one embodiment, the present subject matter includes a methodof analyzing fractions from one or more samples. Each fraction is apart, or portion, of the original sample from which it is obtained orcollected. The original sample can be any material provided for testing,including a biological sample (for example, a pure compound or a mixtureof compounds) wherein the identity, or amount of each component of thesample, is unknown. In one embodiment, the method involves providing oneor more samples to a sampling apparatus that collects successivefractions from each of the samples at discrete points in time. Thediscrete points in time may be equally or unequally spaced from oneanother. In one embodiment, the method involves removing the fractionsfrom the sampling apparatus and then analyzing the fractions with one ormore detector systems that are separate from the sampling apparatus.

[0015] In one embodiment, the present subject matter includes a fractionanalysis system. The system includes an apparatus that collectssuccessive fractions from each of one or more samples at discrete pointsin time. The system also includes one or more detectors, each of whichare separate from the fraction collection apparatus and configured toanalyze the successive fractions.

[0016] In one embodiment, after removal from the fraction collectionapparatus, the collected fractions are available for subsequentprocessing in another process. The present subject matter may beautomated.

[0017] In one embodiment, relevant fractions are combined or isolatedfrom the analytical spectra to produce a purified product on apreparatory scale. Analytical and preparatory modes may be performed onthe same test sample undergoing one pass through the sampling apparatus.

[0018] In one embodiment, the detection system is separate from thecollection system. In one embodiment, the detection system, or detector,is integrated with the collection of fractions.

[0019] In one embodiment, the present subject matter may be used withmultiple detection systems or simultaneously use different detectionsystems. For example, in one embodiment, a CE instrument simultaneouslyprocesses 100 samples, thus producing 100 separate fractionatedcollections, whereby each collection has 384 individual fractions in aspecimen plate. As another example, in one embodiment, the presentsubject matter allows detecting 25 fractionated collections by a firstdetection system (e.g. fluorescence), detecting another 25 fractionatedcollections by a second detection system (e.g. UV-VIS), and detectingthe remaining fractionated collections by a third detection system (e.g.mass spectrometry).

[0020] In one embodiment, a method of testing or analyzing a sampleutilizing continuous sampling techniques enables the direct conversionof analog data into digital signals. The resulting digital datapreserves the analog data and allows analysis (e.g., spectral analysis)at a later time, thus allowing uncoupling of the detector system fromthe sampling apparatus. In one embodiment, an unknown sample iscontinuously analyzed by a method that includes selecting apredetermined time period and waiting for a period of delay. The delayperiod is produced, in part, by latency of migration through the presentsystem. The delay period is determined by the sampling rate. Thesampling rate is selected to be at least twice the highest frequency ofthe smallest discrete moiety present in the unknown sample. Pursuant toNyquist's theorem, the original data is preserved by sampling at twicethe highest frequency. In one embodiment, a sampling rate greater thantwice the highest frequency is used. Successive fractions are collectedat predetermined intervals of time. Fraction collection continues forthe predetermined time period.

[0021] In one embodiment, the present subject matter includes a timesequenced testing apparatus having a sample clock, a sample injector, asampling apparatus, a fraction collector, a computer, and a detector.The sample clock is configured to mark a time period sequence. Thesample injector is adapted to apply one or more samples to a samplingapparatus. The sampling apparatus provides fractions for collecting. Inone embodiment, the sampling apparatus includes a separation pathwaysuch as, for example, a capillary or channel. The fraction collector iscoupled with, and coordinated with the output of the sampling apparatus,and is adapted to receive successive fractions wherein the size andnumber of the fractions are determined by the time period sequence. Thecomputer is adapted to coordinate the sample clock with the fractioncollector and the sampling apparatus. The detector is uncoupled from thesampling apparatus and configured to detect the fractions received fromthe fraction collector after the time period sequence has expired.

[0022] In one embodiment, the apparatus also includes a capillary, acathode electrode, an anode electrode, a power supply, a buffer solutionand a plurality of actuators or movers. The capillary is adapted toperform capillary electrophoresis. The cathode and anode electrodes arepositioned parallel to respective ends of the capillary. The powersupply, adapted for high voltage, is configured to create an electricgradient across the cathode and anode. The buffer solution is comprisedof components non-reactive to the sample. The actuators are adapted tofacilitate transfer of the capillary and electrode from a sample to thebuffer solution, and from the capillary and electrode to the fractioncollector.

[0023] In various embodiments, the present subject matter allowsseparating, identifying, and isolating high volumes of samples usingnanoliter amounts of sample material while limiting human interactionand achieving high resolution. The present subject matter can be usedwith DNA separation, isolation and cloning.

[0024] Other aspects of the invention will be apparent on reading thefollowing detailed description of the invention and viewing the drawingsthat form a part thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

[0025] In the drawings, like numerals describe substantially similarcomponents throughout the several views. Like numerals having differentletter suffixes represent different instances of substantially similarcomponents.

[0026]FIG. 1 illustrates a block diagram of a method in accordance withthe present subject matter.

[0027]FIG. 2 illustrates a schematic diagram of a CE apparatus inaccordance with the present subject matter.

[0028]FIG. 3 illustrates a schematic diagram of a multiple-capillary CEapparatus in accordance with the present subject matter.

DETAILED DESCRIPTION

[0029] In the following detailed description, reference is made to theaccompanying drawings which form a part hereof, and in which is shown byway of illustration specific embodiments in which the invention may bepracticed. These embodiments are described in sufficient detail toenable those skilled in the art to practice the invention, and it is tobe understood that the embodiments may be combined, or that otherembodiments may be utilized and that structural, logical and electricalchanges may be made without departing from the spirit and scope of thepresent invention. The following detailed description is, therefore, notto be taken in a limiting sense, and the scope of the present inventionis defined by the appended claims and their equivalents.

[0030] By way of overview, the present system includes a separationpathway having an input end coupled to a reservoir, or well, of samplematerial. The sample material is migrated through the separation pathwayand fractions are eluted from the output end of the separation pathway.The eluate is received in a collection reservoir or well. The fractionsmove through the separation pathway under a migration field. Themigration field may be created by an electric potential, a pneumaticsource, a vacuum source, or a magnetic source, or other field source.Consider the case of an electric field. In this embodiment, the electricfield is created by an electric potential applied by electrodes incontact with the input end and the output end of the separation pathway.In one embodiment, a first electrode is coupled to the input end of theseparation pathway and a second electrode is coupled to the collectionreservoir. In one embodiment, the collection reservoir includes aplurality of wells, such as for example, a 96-well plate. The secondelectrode is coupled to each well of the 96-well plate. At apredetermined frequency, the output end of the separation pathway isbrought into contact with each successive well of the collectionreservoir, thus setting up an electric field within the pathway.Fractions eluted from the separation pathway migrate into the contactedwell and when the separation pathway is moved away from the collectionreservoir, the migration is halted. A controller coupled to the presentsystem controls the frequency of the contact between the separationpathway and the collection plate. In addition, the controller adjuststhe relative positions to cause each successive fraction to be depositedinto a different well of the collection plate. In this manner, themigratory field is modulated and each well of the collection platereceives a particular fraction eluted at a particular time.

[0031] In one embodiment the controller adjusts the position of theoutput end of the separation pathway and the collection plate remainsstationary. In one embodiment, the separation pathway remains stationaryand the controller adjusts the position of the collection plate. In oneembodiment, both the separation pathway and the collection plate areadjusted by the controller. In one embodiment, the first and secondelectrodes are affixed to the input and output ends, respectively, ofthe separation pathway and the controller modulates the applied voltage.

[0032] In one embodiment, a plurality of separation pathways areprovided with each pathway having an input coupled to a sample well andeach pathway having an output coupled to a multi-well collection plate.For example, in one embodiment, 96 separation pathways are coupled to a96-well input plate and the output of each separation pathway is coupledto a 96-well collection plate. Thus, the output end includes 96collection plates. In one embodiment, each of the 96 collection platesare positioned independently and in another embodiment, each of the 96collection plates are positioned as a group. An actuator coupled to thecollection plate may be coupled to, and operated by, the controller. Theactuator may include an x-axis actuator and a y-axis actuator or arotary actuator. The actuator may also be coupled to the output end ofeach separation pathway.

[0033] The intensity of the migration field, in one embodiment iscontrolled by making, or breaking electrical contact with the collectionplate. In one embodiment, the field intensity is controlled by makingand breaking contact at the input end. Other arrangements are alsocontemplated, such as, for example, a pneumatic system in which appliedair pressure is used to elute fractions from the separation pathway. Inone embodiment, the migration field is provided by a vacuum source. Inone embodiment, a magnetic field, produced by current in electricalwindings in the proximity of, or surrounding, the separation pathway, isenergized to create a migration field. The field magnitude may bemodulated between two intensity levels. For example, in one embodiment,the field magnitude is modulated between zero and a particular uppervalue. As another example, the field magnitude may be modulated betweentwo nonzero values.

[0034] A fraction detector, or detector system, may be positioned at theoutput end of the separation pathway or the collection well. In oneembodiment, fractions are collected without use of a detector andsubsequent processing includes analysis by a detector. In variousembodiments, the detector includes a fluorescent detector, anultraviolet-visible (UV-VIS) detector, a mass spectrometry detector, animmunoassay detector, an electrochemical detector, a radiochemicaldetector, a nuclear magnetic resonance (NMR) detector or a surfaceplasmon resonance (SPR) detector.

[0035] Capillary Electrophoresis Testing Method

[0036]FIG. 1 depicts a testing method 100 practiced according to thepresent invention using CE analysis. It will be appreciated that otherseparation pathways are also contemplated, including for example, amicro-fabricated or nano-fabricated separation pathway. At 110, a timeperiod sequence is defined. Nyquist's theorem for sampling serves as aguide for determining a sampling rate. According to Nyquist the samplingrate must be at least twice the highest frequency of the smallestdiscrete moiety present in the sample in order to reconstruct theoriginal signal. Here, the analog data is preserved digitally bycontinuous sampling at a rate greater than twice the highest frequency.The sampling rate is thus, a function of the time period.

[0037] Consider an example wherein CE is used to separate individualfragments of different size DNA (one base different). The defined timeperiod is determined by choosing a sampling rate that captures no morethat one base pair per sampling. Thus, the defined time period capturesthe smallest discrete moiety in the sampling. The summation of all thesetime periods over the entire time a sample is analyzed constitutes thetime period sequence. The time period sequence may include a finitenumber of equally spaced time periods. In one embodiment, the timeperiods differ logarithmically, exponentially, or geometrically. In oneembodiment, the sequence of time periods is experimentally determined.The time period sequence may be defined by any method known in the artof continuous sampling.

[0038] At 115, a test sample is introduced into the CE instrument. Inone embodiment, the test sample is injected, however, other methods ofapplying the sample to the CE instrument are also contemplated. Thesample may be robotically or manually introduced. It will also beappreciated that other analytical or preparatory devices are alsocontemplated. For example, immunoassay, or high performance liquidchromatography (HPLC), or other assay techniques may also be used. Thesample may include a mixture of proteins, macromolecules, nucleotides,carbohydrates, enantiomers, small molecule libraries or naturalcompounds.

[0039] At 120, a voltage is applied across the CE capillary. In oneembodiment, the medium within the capillary, or separation pathway, andthe characteristics of the test sample determine the voltage applied.

[0040] At 125, a time period elapses. The time period is determined bythe time period sequence at 110. During this time period, an electricgradient exists across the separation pathway due to the voltage appliedat 120. The gradient resolves and separates the individual components inthe test sample. In one embodiment, a two hour time period isestablished and fraction collection occurs every 30 seconds after aninitial delay period of one hour.

[0041] At 130, the voltage from the capillary is removed followingexpiration of the time period at 125. Thus, the present subject matterachieves continuous sampling. In one embodiment, sampling does not occurafter removal of the voltage from the capillary and the electricgradient is removed. Thus, a fraction is captured when the voltage isapplied.

[0042] At 135, a fraction is collected corresponding to the time periodduring which the electric gradient exists across the capillary. Thecollected fraction includes the material collected during the timeperiod in which analysis occurs. In one embodiment, the fraction iscollected in an individual well of a standard specimen plate, forexample, a 96-well or 384-well specimen plate. The fraction may bemanually or robotically collected. Other devices used to hold fractionsare contemplated within the present invention. For example, test tubes,blotting paper, or individual vials may be used.

[0043] In one embodiment, after collecting a fraction at 135, thespecimen plate is moved into position to receive the next fraction, at140. For example, the specimen plate may be moved robotically. In oneembodiment, the separation pathway, or capillary, is moved to manipulatethe output into the next well of the specimen plate. In one embodiment,the methods from 120 through 140 are repeated through each successivetime period 125 until the last time period expires in the time periodsequence defined at 110. The method collects fractions when an electricgradient is applied across the capillary, thus ensuring continuoussampling of the test sample throughout the entire analysis.Consequently, each fraction has been captured within a discrete timeperiod on the specimen plate. In one embodiment, the sampling time issynchronized with the mobility change of the analyte.

[0044] After the last time period, at 150, the last fraction iscollected, at 155. In one embodiment, at 160, the specimen plate isremoved from the CE instrument. After removal from the CE instrument, at165, the contents of the specimen plate are detected. In one embodiment,detection includes, for example, charge-coupled device (CCD) arraysusing an ultraviolet (UV) or fluorescence monitor may detect the entirespecimen plate at one time. Alternatively, the specimen plate may bedetected individually or row by row. In one embodiment, the specimenplate undergoes more than one detection process. For example, thespecimen plate may be monitored first by V, then fluorescence, and thenby mass spectrometry. Other detection modes, such as conductivity,electrochemical, or radioactive means are also contemplated. In oneembodiment, the detector includes a fluorescent detector, anultraviolet-visible (UV-VIS) detector, a mass spectrometry detector, animmunoassay detector, an electrochemical detector, a radiochemicaldetector, a nuclear magnetic resonance (NMR) detector or a surfaceplasmon resonance (SPR) detector.

[0045] In one embodiment, the method according to FIG. 1 is practicedusing a CE instrument that separates individual base pairs of DNA. Oncea detector detects the fractions in the specimen plate, a spectra isproduced of the separation in which each individual peak corresponds toan individual base pair. From this analytical spectra, desired basepairs may be isolated and the corresponding fraction amplified bypolymerase chain reaction (PCR), thus creating preparative amounts ofisolated and purified base pairs.

[0046] In one embodiment, the CE analysis may be automated. For example,detection, at 165, may be accomplished using a high throughput system.Further, creating preparatory amounts of a specific fraction in thespecimen plate may also occur robotically using a high throughputsystem. Accordingly, the present subject matter provides for anautomated process of testing large numbers of samples in a highthroughput system. In one embodiment, detection is uncoupled from thesampling apparatus and both analytical and preparatory modes may bepracticed on a single pass though the sampling apparatus. More than onedetection device may be utilized on the same specimen plate. In thisway, high volumes of samples may be analyzed using multiple detectionsystems in both an analytical and preparatory mode using a small amountof material.

[0047] Capillary Electrophoresis Sampling Apparatus

[0048] In accordance with the present invention, a diagram of system 200is provided in FIG. 2. In the embodiment shown, sampling apparatus 200includes a CE instrument used to separate, isolate, and resolve mixturesof proteins, macromolecules, nucleotides, enantiomers, and chiralmolecules based on the differences in molecular charged to mass ratios.In this embodiment, the CE instrument is configured to sequence DNAfragments and isolate individual base pairs.

[0049] In one embodiment, capillary 210 is filled with a molecularsieving matrix, such as polyacrylamide, polyethylene oxide or othertypes of polymers. Other types of gel may also be used. It will also beappreciated that other CE techniques such as isoelectric focusing,isotachophoresis (ITP), and hydrophrobicity (micellar electrokineticcapillary chromatography, MECC), and other CE techniques may be used.Coupled to capillary 210 is electrode 225. Electrode 225 includes anode215 at one end and cathode 220 at the other end of the capillary. Bymeans of power supply 230, a high voltage is applied across electrode225, creating a positive charge at anode 215 and a negative charge atcathode 220. This in turn creates an electric gradient across capillary210. Voltmeter 235 is connected to power supply 230 and indicates thevoltage applied to electrode 225.

[0050] In one embodiment, electrode 225 and capillary 210 are positionedinside sample reservoir 255 holding test sample 260. Injector 250coordinates injection of test sample 260 into capillary 210. Otherinjectors 250 and sample holders 255 may be used to apply sample 260 tocapillary 210. For example, an automated injector system in which thesample holder 255 includes a syringe may also be used.

[0051] In one embodiment, sample holder 255 includes an individual wellin a 96-well, or larger or smaller, specimen plate.

[0052] Vertical sample mover 270 and horizontal sample mover 275, (alsoreferred to as actuators) are represented in the figure by directionalarrows. In one embodiment, mover 270 and mover 275 are positioned tomove sample holder 255, buffer holder 263, or capillary 210, such thatcapillary 210 and electrode 225 are in contact with the contents ofreservoir 255 or 263. For example, vertical and horizontal movers 270and 275 are operable to move sample container 255 out of contact withcapillary 210 and electrode 225 after the test sample is injected.Movers 275 and 270 position buffer container 263 such that buffersolution 265 is in contact with electrode 225 and capillary 210. Movers270 and 275 may be manual or robotic. In one embodiment, the actuatorsinclude one or more linear or rotary motors.

[0053] In one embodiment, computer 240 coordinates the actions ofinjector 250, power supply 230, volt meter 235, and time period clock245, and movers 270, 275, 285 and 280. Computer 240 executes a computerprogram to coordinate the electric field gradient intensity with thecollecting of fractions. In one embodiment, system 200 is configuredsuch that injector 250 applies sample 260 to capillary 210. Movers 270and 275 position buffer container 263 such that capillary 210 andelectrode 225 is immersed in buffer solution 265. In this embodiment,the anodic end of electrode 225 is immersed in buffer solution 265.

[0054] Computer 240, in one embodiment, includes a processor withmemory, a user input device (such as a keyboard or mouse), an outputdevice (such as a display or printer). The memory contents can includeprogram memory or data derived from the present subject matter.

[0055] In one embodiment, computer 240 instructs power supply 230 toapply a voltage across electrode 225 such that the anodic end 215 of theelectrode carries a positive charge while the cathodic end of theelectrode 220 carries a negative charge. Thus, an electric gradientforms across capillary tube 210. The electric gradient is maintainedacross electrode 210 for a defined time period marked by clock 245.After the time period marked by clock 245 expires, the voltage suppliedby power supply 230 is removed, thereby removing the electric gradientacross capillary 210. Once the electric field is removed, analysis by CEis suspended or interrupted. Interruption of the migration field mayinclude terminating the field or modulating the field between two ormore non-zero intensity levels.

[0056] During the time period, buffer solution is drawn up from buffercontainer 263 and drawn through capillary 210 and collected in fractioncollector plate 290 in an individual collector well 295. In oneembodiment, fraction collection occurs while the voltage is appliedacross electrode 225. Fraction collector plate 290 may include, forexample, a 96-well specimen plate, an array of vials, or other specimenplates.

[0057] After the time period marked by clock 245 expires and powersupply 230 has removed the voltage across electrode 225, vertical mover280 and horizontal mover 285 position fraction collector plate 290 suchthat a next individual collector well 295 is positioned to receive thenext fraction from capillary 210. After fraction collector plate 290 ispositioned to receive the next fraction from capillary 210, clock 245begins measuring a successive time period triggering application of avoltage across electrode 225 supplied by power supply 230. During thistime period, the next fraction is collected from capillary 210 by thesuccessive individual fraction well 295.

[0058] The time periods marked by clock 245 may be uniform or differentfor each successive time period. For example, each time period measuredmay be 30 seconds in duration, or, alternatively, the first time periodmay be 90 seconds to account for the void volume of the capillary 210,and successive fractions may be collected on a 30 second basis. Asanother example, time periods may be measured logarithmically,geometrically or exponentially. In one embodiment, the sampling time issynchronized with the mobility change of the analyte. For example, wheremobility of an analyte is half as fast, the time period is twice aslong.

[0059] In one embodiment, after sampling is complete, movers 280 and 285transport the fraction collection plate 290 to a detection processingarea. A second sample may be analyzed while the first sample is beingdetected at another processing station. In one embodiment, each fractioncollector plate undergoes multiple detection methods after removal fromsystem 200.

[0060] Multiple-Capillary and Capillary Electrophoresis Apparatus

[0061] System 300 in accordance with one embodiment of the presentsubject matter is illustrated in FIG. 3. In this embodiment, system 300includes multiple capillaries by which multiple test samples aresimultaneously analyzed.

[0062] The embodiment illustrated in FIG. 3 employs an array ofcapillaries 330 or separation pathways. The separation pathways mayincluding a plurality of individual capillaries 210 which may includemicrofabricated or nanofabricated channels. A corresponding array ofelectrodes 335, including individual electrodes 225, are coupled tocapillary array 330 such that each electrode 225 is coupled to acorresponding capillary 210. Each individual capillary 210 and itscorresponding electrode 225 is in contact with an individual test samplewell 315. Collectively, these individual test sample wells 315 form anarray of sample wells 310. In one embodiment, this array of sample wells310, in which each individual sample well 315 contains a test sample260, may be a 96-well sample plate. Other sample arrays are alsocontemplated. For example, a collection of vials or test tubes may beused.

[0063] Each test sample 260 contained in individual sample well 315 maybe identical to other test samples contained in sample array 310 or thetest samples may vary across the array. For example, each individualsample well within the array of sample wells may contain a different DNAfragment to be sequenced. Conversely, non-redundant, expressed sequencetag (EST) libraries may be constructed used in connection with otherhigh throughput processes.

[0064] The 96-well specimen plate does not limit the number ofcapillaries that may be used in this apparatus at any one time. Forexample, a 384-well sample plate may be used in which 384 capillariesand 384 varied or identical samples may be simultaneously analyzed.Fractions for each capillary 210 are collected in fraction collectorplate array 390, wherein array 390 includes a plurality of fractioncollector plates 290 configured to receive fractions. Each individualcapillary 210 corresponds to an individual fraction collection plate290. Movers 280 and 285 coordinate positioning of the fraction collectorplates relative to the capillaries, to receive successive fractions.

[0065] In one embodiment, multiple samples are analyzed and detected ata subsequent processing station. For example, detection may occur aspart of a high throughput system such as a CCD array configured forultraviolet-visible (UV-VIS) or fluorescence detection. In oneembodiment, multiple detection systems are used. For example, somefractions may undergo UV-VIS detection while other fractions undergofluorescence detection and still other fractions undergo both UV-VIS andfluorescent detection.

[0066] The present invention may be practiced in both an analytical modeand a preparatory mode. In one embodiment, a sample undergoes CEanalysis, thus creating multiple fractions on a specimen plate. At alater time, the specimen plate may be detected using laser-inducedfluorescence, thus generating an analytical spectra of the processedsample. From this spectra, certain peaks corresponding to certainfractions may be amplified and duplicated, for example, using PCR orcloning. In this manner, preparatory amounts of certain fractions havebeen generated from the same specimen plate that provided the analyticaldata.

[0067] In the figure, plate 310 is shown coupled electrically to powersupply 230 by electrode 215. In addition, array 390 is shown coupledelectrically to power supply 230 by electrode 220. Each plate 290 withinarray 390 is coupled electrically to electrode 220. In the embodimentshown, plates 310, 290 and array 390 are electrically conductive, andfabricated of such materials as a metal or conductive ceramic. In oneembodiment, plate 310 is fabricated of non-conductive, orsemiconductive, material and each well, or reservoir, 315 is lined withan electrically insulative material and electrode 210 is coupled to eachwell 315 by an individual electrode. In one embodiment, array 390, orplates 290, are fabricated of non-conductive, or semiconductive,material and each well or reservoir 295 in plate 290 is lined with anelectrically insulative material and electrode 220 is coupled to eachwell 295 by an individual electrode.

[0068] Any number of fractions may be collected without regard tocorrelating a detected peak to a specific fraction during the analysis.

[0069] Alternate Embodiments

[0070] In one embodiment, each separation pathway is associated with aparticular collection plate having a plurality of collection wells.Thus, 96 collection plates are used in a system having 96 separationpathways. In this manner, each separation pathway is individuallycontrollable relative to the associated collection plate for thatpathway. In one embodiment, the separation pathway is stationary and thecollection plate is positionable by an actuator. The collection platesare mounted in a frame or otherwise synchronized to move together. Inone embodiment, the collection plates are stationary and the separationpathways are positionable by an actuator. In one embodiment, eachseparation pathway is positioned independently of the position of otherpathways. In one embodiment, each collection plate is positionedindependently of the position of other plates. In one embodiment, oneactuator, or set of actuators, controls movement of a collection plate(or array of collection plates) along a first axis, such as an x-axis. Asecond actuator, or set of actuators, controls movement of a separationpathway along a second axis, such as a y-axis. Other arrangements ofactuators are also contemplated.

[0071] The actuators may include one or more linear or rotary actuators,or motors. For example a first linear motor controls movement of acollection plate along an x-axis and a second linear motor controlsmovement of the plate along a y-axis. Rotary actuators may also be usedto control the relative position of the separation pathway relative tothe collection plate. The actuators may include a pneumatic cylinder, alead screw, a hydraulic cylinder, an electric solenoid or a magneticactuator.

[0072] Conclusion

[0073] The above-described system provides, among other things, asystem, apparatus and method for collection and analysis with highresolution and high throughput.

[0074] It will be appreciated that the methods described herein may beperformed in different orders than described and that portions of amethod may be repeated.

[0075] It is to be understood that the above description is intended tobe illustrative, and not restrictive. Many other embodiments will beapparent to those of skill in the art upon reviewing the abovedescription. The scope of the invention should, therefore, be determinedwith reference to the appended claims, along with the full scope ofequivalents to which such claims are entitled.

What is claimed is:
 1. A system comprising: a separation pathway havinga first end and a second end; a sample well in communication with thefirst end; one or more collection wells, wherein the second end isadapted to communicate with at least one collection well of the one ormore collection wells; a power supply having a first electrode and asecond electrode adapted to create an electric field between the firstend and the second end; a first actuator adapted to adjust a firstposition of the second end relative to the plurality of collectionwells; and a controller coupled to the first actuator and adapted tomodulate a potential between the first end and the second end andadapted to control the first position.
 2. The system of claim 1 furthercomprising a detector in communication with the second end.
 3. Thesystem of claim 2 wherein the detector includes a fluorescent detector,an ultraviolet-visible (UV-VIS) detector, a mass spectrometry detector,an immunoassay detector, an electrochemical detector, a radiochemicaldetector, a nuclear magnetic resonance (NMR) detector or a surfaceplasmon resonance (SPR) detector.
 4. The system of claim 1 wherein thefirst electrode is coupled to the first end.
 5. The system of claim 1wherein the first electrode is coupled to the sample well.
 6. The systemof claim 1 wherein the second electrode is coupled to the second end. 7.The system of claim 1 wherein the second electrode is coupled to the oneor more collection wells.
 8. The system of claim 1 wherein theseparation pathway includes a capillary or microchannel.
 9. The systemof claim 1 wherein the actuator includes a motor coupled to theplurality of collection wells.
 10. The system of claim 1 wherein thefirst actuator includes a first motor adapted to displace the pluralityof collection wells along a first axis and a second motor adapted todisplace the plurality of collection wells along a second axis.
 11. Thesystem of claim 1 wherein the first actuator includes a motor coupled tothe second end.
 12. The system of claim 1 wherein the first actuatorincludes a first motor adapted to displace the second end along a firstaxis and a second motor adapted to displace the second end along asecond axis.
 13. The system of claim 1 wherein the controller includes aprocessor.
 14. The system of claim 1 further including: a plurality ofsample wells wherein the sample well in communication with the first endincludes a selected sample well of the plurality of sample wells; asecond actuator adapted to adjust a second position of the first endrelative to the plurality of sample wells; and wherein the controller isadapted to control the second position.
 15. The system of claim 1wherein the second actuator includes a motor coupled to the plurality ofsample wells.
 16. The system of claim 1 wherein the second actuatorincludes a third motor adapted to displace the plurality of sample wellsalong a first axis and a fourth motor adapted to displace the pluralityof sample wells along a second axis.
 17. The system of claim 1 whereinthe second actuator includes a motor coupled to the first end.
 18. Thesystem of claim 1 wherein the second actuator includes a third motoradapted to displace the first end along a first axis and a fourth motoradapted to displace the first end along a second axis.
 19. The system ofclaim 1 wherein the controller includes a clock.
 20. The system of claim1 wherein the controller includes a voltage controller coupled to thepower supply.
 21. A computer implemented method comprising: applying asample to an input of a separation pathway; generating a migratory fieldin the separation pathway; eluting an analyte of the sample from theseparation pathway; collecting the analyte in a collection well;interrupting the migratory field after collecting commences; andrepeating the collecting and the interrupting, at a predetermined timeinterval, for a successive analtye and a successive collection well. 22.The method of claim 21 wherein repeating the collecting andinterrupting, at the predetermined time interval includes repeating thecollecting and interrupting, at substantially uniformly spaced timeintervals.
 23. The method of claim 21 further comprising synchronizingthe collecting and interrupting with the mobility of the analtye. 24.The method of claim 21 further comprising analyzing the analtye prior tocollecting.
 25. The method of claim 21 wherein injecting the sampleincludes injecting a biological sample.
 26. The method of claim 21wherein injecting a sample includes injecting a mixture of proteins,macromolecules, nucleotides, carbohydrates, enantiomers, small moleculelibraries or natural compounds.
 27. The method of claim 21 whereincreating a migratory field includes applying a potential to theseparation pathway.
 28. The method of claim 21 wherein creating amigratory field includes applying a pressure to the separation pathway.29. The method of claim 21 wherein creating a migratory field includesdrawing a vacuum in the separation pathway.
 30. The method of claim 21wherein collecting includes positioning the separation pathway relativeto the collection well.
 31. The method of claim 21 wherein repeatedlyinterrupting the migratory field includes adjusting a potential withinthe separation pathway.
 32. The method of claim 21 further comprisingestablishing the predetermined time interval as a function of acomposition of the separation pathway.
 33. A system comprising: aplurality of separation pathways, each separation pathway having a firstend and a second end; a plurality of sample wells, wherein each samplewell is in communication with a first end of a separation pathway; apower supply having a first electrode and a second electrode adapted tocreate an electric field between the first end and the second end ofeach separation pathway; for each separation pathway, a plurality ofcollection wells wherein each collection well is adapted to communicatewith a second end of the separation pathway; for each separationpathway, a first actuator adapted to adjust a position of the second endrelative to the plurality of collection wells; and a controller coupledto the first actuator and adapted to modulate the electric field andadapted to control the position.
 34. The system of claim 33 furthercomprising a detector coupled to each separation pathway of theplurality of separation pathways.
 35. The system of claim 33 furthercomprising a detector coupled to each collection well of the pluralityof collection wells.
 36. The system of claim 33 wherein the plurality ofseparation pathways includes a multichannel capillary.
 37. The system ofclaim 33 wherein the plurality of separation pathways includes aplurality of microchannel pathways.
 38. The system of claim 33 whereinthe plurality of separation pathways includes a plurality of nanochannelpathways.
 39. The system of claim 33 wherein the plurality of samplewells includes a multi-well plate.
 40. The system of claim 33 whereinthe plurality of collection wells includes a multi-well plate.
 41. Thesystem of claim 33 further comprising a frame wherein each of theplurality of collection wells for each separation pathway is secured tothe frame.
 42. The system of claim 40 wherein the first actuator iscoupled to the frame.
 43. The system of claim 33 wherein the firstactuator is coupled to the plurality of separation pathways.
 44. Thesystem of claim 33 wherein the first actuator is coupled to theplurality of collection wells.